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Related Experiment Video

Updated: Mar 26, 2026

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions

Published on: April 4, 2014

Fragment-Based Screening for Enzyme Inhibitors Using Calorimetry.

Michael I Recht¹, Vicki Nienaber², Francisco E Torres¹

¹Palo Alto Research Center, Palo Alto, California, USA.

Methods in Enzymology

|January 23, 2016

Summary

This summary is machine-generated.

Isothermal titration calorimetry (ITC) offers sensitive, label-free analysis of binding and enzyme reactions. Nanocalorimetry enhances conventional ITC for efficient screening of compound libraries, overcoming limitations in reagent use and time.

Keywords:

Enzyme assay Fragment library Fragment-based lead discovery Label-free assay Microcalorimetry Nanocalorimetry

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Last Updated: Mar 26, 2026

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Area of Science:

Biophysical Chemistry
Enzyme Kinetics
Drug Discovery

Background:

Isothermal titration calorimetry (ITC) is a sensitive method for studying binding thermodynamics and label-free enzyme kinetics.
Conventional microcalorimetry is valuable for hit validation but limited in primary screening due to high reagent requirements and long assay times.

Purpose of the Study:

To describe the application of nanocalorimetry and conventional microcalorimetry for screening compound libraries.
To highlight nanocalorimetry's potential to overcome limitations of conventional ITC in fragment-based lead discovery.

Main Methods:

Utilized isothermal titration calorimetry (ITC) with both conventional microcalorimeters and advanced nanocalorimeters.
Applied label-free measurement techniques for enzymatic reactions.
Screened compound libraries, including those typical in fragment-based lead discovery.

Main Results:

Demonstrated the utility of nanocalorimetry for screening large compound libraries (500-1000 compounds).
Showcased nanocalorimetry's ability to overcome the limitations of conventional ITC regarding reagent consumption and measurement duration.
Validated the use of microcalorimetry for hit validation and characterization.

Conclusions:

Nanocalorimetry offers a viable solution for primary screening of compound libraries in drug discovery.
Both nanocalorimetry and conventional microcalorimetry are powerful tools for enzyme inhibitor screening.
Label-free calorimetric methods provide essential thermodynamic and kinetic data for characterizing molecular interactions.